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Development of a DNA vaccine against chicken anemia virus by using a bicistronic vector expressing VP1 and VP2 proteins of CAV

Moeini H, Omar AR, Rahim RA, Yusoff K

Comp Immunol Microbiol Infect Dis, 2011 May;34(3):227-36.
PMID: 21146874 DOI: 10.1016/j.cimid.2010.11.006

In the present study, we describe the development of a DNA vaccine against chicken anemia virus. The VP1 and VP2 genes of CAV were amplified and cloned into pBudCE4.1 to construct two DNA vaccines, namely, pBudVP1 and pBudVP2-VP1. In vitro and in vivo studies showed that co-expression of VP1 with VP2 are required to induce significant levels of antibody against CAV. Subsequently, the vaccines were tested in 2-week-old SPF chickens. Chickens immunized with the DNA-plasmid pBudVP2-VP1 showed positive neutralizing antibody titer against CAV. Furthermore, VP1-specific proliferation induction of splenocytes and also high serum levels of Th1 cytokines, IL-2 and IFN-γ were detected in the pBudVP2-VP1-vaccinated chickens. These results suggest that the recombinant DNA plasmid co-expressing VP1 and VP2 can be used as a potential DNA vaccine against CAV.
New sequential treatment for mature landfill leachate by cationic/anionic and anionic/cationic processes: optimization and comparative study

Bashir MJ, Aziz HA, Yusoff MS

J Hazard Mater, 2011 Feb 15;186(1):92-102.
PMID: 21146925 DOI: 10.1016/j.jhazmat.2010.10.082

Two new applications for sequence treatment of mature (stabilized) landfill leachate, that is, cationic resin followed by anionic resin (cationic/anionic) and anionic resin followed by cationic resin (anionic/cationic), are employed and documented for the first time in the literature. Response surface methodology (RSM) concerning central composite design (CCD) is used to optimize each treatment process, as well as evaluate the individual and interactive effects of operational cationic resin dosage and anionic resin dosage on the effectiveness of each application in terms of color, chemical oxygen demand (COD), and NH(3)-N removal efficiency. A statistically significant model for color, COD, and NH(3)-N removal was obtained with high coefficient of determination values (R(2)>0.8). Under optimum operational conditions, the removal efficiency levels for color, COD, and NH(3)-N are 96.8%, 87.9%, and 93.8% via cationic/anionic sequence, and 91.6%, 72.3%, and 92.5% via anionic/cationic sequence, respectively. The experimental results and the model predictions agree well with each other.
Fever/clinical signs and association with neutropenia in solid cancer patients--bacterial infection as the main cause

Rasool Hassan BA, Yusoff ZB, Othman SB

Asian Pac J Cancer Prev, 2010;11(5):1273-7.
PMID: 21198276

INTRODUCTION: Neutropenia remains one of the serious side effects of chemotherapeutics drugs making cancer patients face serious risk of infections. Fever and clinical signs are considered as important indicators. The objectives of this study were to assess fever and clinical signs with neutropenia onset and/ or severity in solid cancer cases, using culture tests to determine the type of bacteria predominating, whether gram positive or gram negative.
METHODS: This observational retrospective study was conducted on files of all solid cancer patients who admitted to a general hospital between 1 January 2003 and 31 December 2006. All data were categorical and analyzed for association with neutropenia.
RESULTS: 117 neutropenic patients were studied, 83 (70.9%) of them suffering from fever ranging between 38.5-39 °C, with hypotension (53; 27.3%) and headache 51 (26.3%) as the most common clinical signs. Only 34 (29.1%) neutropenic patients underwent culture testing and only 14 (41.2%) showed positive growth, gram negative types predominating (9; 64.2%), mainly Escherichia coli (5; 35.7%), with gram positive only in 5 (35.7%). Significant associations were found for fever and clinical signs with neutropenia severity (P<0.05), but not neutropenia onset (P>0.05). Logistic regression results showed strong significant association between presence of fever (P=0.02, OR=1.3) (95% confidence interval (CI)) hypotension and headache (P=0.001, OR=1.148) (95% CI) with neutropenia severity.
CONCLUSION: Fever and clinical signs specifically headache and hypotension are symptoms associated with severe neutropenia in solid cancer patients. Both may primarily result from bacterial infection, particularly gram negative forms.
Fulltext Role of excitatory amino acid input in rostral ventrolateral medulla neurons in rats with obesity-induced hypertension

Suhaimi FW, Yusoff NH, Dewa A, Yusof AP

Acta Neurol Belg, 2010 Mar;110(1):57-64.
PMID: 20514927

Obesity is intimately associated with hypertension; increases in blood pressure are closely related to the magnitude of weight gain. The present study aims to determine whether the excitatory amino acid input to rostral ventrolateral medulla (RVLM) contributes to elevated blood pressure in rats with diet-induced obesity. Male Sprague-Dawley rats weighing 280 to 300 grams were fed with a low-fat diet (10% kcal from fat) or moderately high-fat diet (32% kcal from fat) for 16 weeks. At week 16, rats on the moderate high-fat diet were segregated into obesity-prone and obesity-resistant rats based on body weight distribution. Baseline mean arterial pressure (MAP) was significantly higher in obesity-prone rats as compared to obesity-resistant and rats on a low-fat diet. Bilateral injection of kynurenic acid (KYN) (40 nM) into the RVLM of the obesity-prone rats reduced MAP to levels significantly different from those observed in rats on a low-fat diet and obesity-resistant rats (no change in MAP). At a lower concentration (4 nM), KYN injection did not produce any change in MAP in any group. The results obtained suggest that excitatory amino acid input to the RVLM does contribute to the development of hypertension in rats with diet-induced obesity.
Fulltext Improving the potency of DNA vaccine against chicken anemia virus (CAV) by fusing VP1 protein of CAV to Marek's Disease Virus (MDV) type-1 VP22 protein

Moeini H, Omar AR, Rahim RA, Yusoff K

Virol J, 2011;8:119.
PMID: 21401953 DOI: 10.1186/1743-422X-8-119

Studies have shown that the VP22 gene of Marek's Disease Virus type-1 (MDV-1) has the property of movement between cells from the original cell of expression into the neighboring cells. The ability to facilitate the spreading of the linked proteins was used to improve the potency of the constructed DNA vaccines against chicken anemia virus (CAV).
Ellagic acid, phenolic acids, and flavonoids in Malaysian honey extracts demonstrate in vitro anti-inflammatory activity

Kassim M, Achoui M, Mustafa MR, Mohd MA, Yusoff KM

Nutr Res, 2010 Sep;30(9):650-9.
PMID: 20934607 DOI: 10.1016/j.nutres.2010.08.008

Natural honey has been used in traditional medicine of different cultures throughout the world. This study looked into the extraction of Malaysian honey and the evaluation of the anti-inflammatory activity of these extracts. It was hypothesized that honey extracts contain varying amounts of phenolic compounds and that they possess different in vitro anti-inflammatory activities. Honey extracts were analyzed using liquid chromatography-mass spectrometry to identify and compare phenolic compounds, whereas high-performance liquid chromatography was used for their quantification. Subsequently, honey methanol extract (HME) and honey ethyl acetate extract (HEAE) were tested in vitro for their effect on nitric oxide production in stimulated macrophages. The extracts were also tested for their effects on tumor necrosis factor-α (TNF) cytotoxicity in L929 cells. The major phenolics in the extracts were ellagic, gallic, and ferulic acids; myricetin; chlorogenic acid; and caffeic acid. Other compounds found in lower concentrations were hesperetin, p-coumaric acid, chrysin, quercetin, luteolin, and kaempferol. Ellagic acid was the most abundant of the phenolic compounds recorded, with mean concentrations of 3295.83 and 626.74 μg/100 g of honey in HME and HEAE, respectively. The median maximal effective concentrations for in vitro nitric oxide inhibition by HEAE and HME were calculated to be 37.5 and 271.7 μg/mL, respectively. The median maximal effective concentrations for protection from TNF cytotoxicity by HEAE and HME were 168.1 and 235.4 μg/mL, respectively. In conclusion, HEAE exhibited greater activity in vitro, whereas HME contained a higher concentration of phenolic compounds per 100 g of honey.
Lipid biosynthesis in Cunninghamella bainieri 2A1 in N-limited and N-excess media

Taha EM, Omar O, Yusoff WM, Hamid AA

Annals of microbiology, 2010 Dec;60(4):615-622.
PMID: 21125005

Lipid biosynthesis and fatty acids composition of oleaginous zygomycetes, namely Cunninghamella bainieri 2A1, cultured in media with excess or limited nitrogen were quantitatively determined at different times of culture growth. Accumulation of lipids occurred even when the activity of NAD(+)-ICDH (β-Nicotinamide adenine dinucleotide-isocitrate dehydrogenase) was still detectable in both media. In C. bainieri 2A1, under nitrogen limitation, the ratio of lipids was around 35%, whereas in nitrogen excess medium (feeding media supplemented with ammonium tartarate), the lipid ratio decreased. The amount of this decrease depended on the level of ammonium tartarate in the media. The main findings in this paper were that C. bainieri 2A1 has the ability to accumulate lipid although nitrogen concentration detected inside the media and that NAD-ICDH was active in all culture periods. These results proved that the strain C. bainieri 2A1 has an alternative behavior in lipid biosynthesis that differs from yeast. According to the old hypotheses, yeasts could not accumulate lipid more than 10% when nitrogen was detected inside the media. Nitrogen-limited and excess media both contained the same fatty acids (palmitic acid, stearic acid, olic acid, linoleic acid and γ-linolenic acid), but at different concentrations. The C:N ratio was also studied and showed no effects on total lipid accumulation, but a significant effect on γ-linolenic acid concentration.
Role of supraspinal vasopressin neurones in the effects of atrial natriuretic peptide on sympathetic nerve activity

Yusof AP, Yusoff NH, Suhaimi FW, Coote JH

Auton Neurosci, 2009 Jun 15;148(1-2):50-4.
PMID: 19349212 DOI: 10.1016/j.autneu.2009.03.005

The aim of the present study was to determine if paraventricular-spinal vasopressin neurones participate in the sympatho-inhibitory effects of systemically administered atrial natriuretic peptide (ANP) on renal sympathetic nerve activity (RSNA). Experiments were carried out on male Sprague-Dawley rats anesthetized with 1.3 g/kg urethane. Changes in mean arterial pressure (mm Hg), heart rate (beats per minute) and RSNA (%) were measured following intravenous bolus administration of ANP (250 ng, 500 ng and 5 microg). Intrathecal application of selective V 1a receptor antagonist was performed to test for the involvement of supraspinal vasopressin pathways in mediating the effect on sympathetic outflow evoked by intravenous ANP administration. The results obtained demonstrated that both low and high doses of ANP caused renal sympathoinhibition (250 ng; - 7.5 +/- 1%, 500 ng; - 14.2 +/- 1%, 5 microg; - 16.4 +/- 2%), concomitant with vasodilation and bradycardia. After spinal vasopressin receptor blockade, the inhibitory effects of ANP were prevented and there was a small renal sympatho-excitation (250 ng; + 1.7 +/- 0.2%, 500 ng; + 6.1 +/- 0.03%, 5 microg; + 8.0 +/- 0.03%, P < 0.05). Therefore, the renal sympathetic nerve inhibition elicited by circulating ANP is dependent on the efficacy of a well established supraspinal vasopressin pathway. Since supraspinal vasopressin neurones without exception excite renal sympathetic neurones, it is suggested that ANP elicits this effect by activating cardiac vagal afferents that inhibit the spinally projecting vasopressin neurones at their origin in the paraventricular nucleus of the hypothalamus.
DNA vaccine encoding avian influenza virus H5 and Esat-6 of Mycobacterium tuberculosis improved antibody responses against AIV in chickens

Oveissi S, Omar AR, Yusoff K, Jahanshiri F, Hassan SS

Comp Immunol Microbiol Infect Dis, 2010 Dec;33(6):491-503.
PMID: 19781778 DOI: 10.1016/j.cimid.2009.08.004

The H5 gene of avian influenza virus (AIV) strain A/chicken/Malaysia/5744/2004(H5N1) was cloned into pcDNA3.1 vector, and Esat-6 gene of Mycobacterium tuberculosis was fused into downstream of the H5 gene as a genetic adjuvant for DNA vaccine candidates. The antibody level against AIV was measured using enzyme-linked immunosorbent assay (ELISA) and haemagglutination inhibition (HI) test. Sera obtained from specific-pathogen-free chickens immunized with pcDNA3.1/H5 and pcDNA3.1/H5/Esat-6 demonstrated antibody responses as early as 2 weeks after the first immunization. Furthermore, the overall HI antibody titer in chickens immunized with pcDNA3.1/H5/Esat-6 was higher compared to the chickens immunized with pcDNA3.1/H5 (p<0.05). The results suggested that Esat-6 gene of M. tuberculosis is a potential genetic adjuvant for the development of effective H5 DNA vaccine in chickens.
Association of ABO blood group B with myocardial infarction

Sheikh MK, Bahari MB, Yusoff NM, Knight A

J Coll Physicians Surg Pak, 2009 Aug;19(8):514-7.
PMID: 19651016 DOI: 08.2009/JCPSP.514517

The purpose of this study was to find out the association between blood group B and Myocardial Infarction (MI) in sample population in Malaysia.
Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmid-Mg/Al-LDH nanobiocomposite material as a vector for gene expression

Masarudin MJ, Yusoff K, Rahim RA, Hussein MZ

Nanotechnology, 2009 Jan 28;20(4):045602.
PMID: 19417322 DOI: 10.1088/0957-4484/20/4/045602

The delivery of a full plasmid, encoding the green fluorescent protein gene into African monkey kidney (Vero3) cells, was successfully achieved using nanobiocomposites based on layered double hydroxides. This demonstrated the potential of using the system as an alternative DNA delivery vector. Intercalation of the circular plasmid DNA, pEGFP-N2, into Mg/Al-NO(3)(-) layered double hydroxides (LDH) was accomplished through anion exchange routes to form the nanobiocomposite material. The host was previously synthesized at the Mg(2+) to Al(3+) molar ratio R(i) = 2 and subsequently intercalated with plasmid DNA. Size expansion of the interlamellae host from 8.8 A in LDH to 42 A was observed in the resulting nanobiocomposite, indicating stable hybridization of the plasmid DNA. The powder x-ray diffraction (PXRD) results, supplemented with Fourier-transform infrared (FTIR) spectroscopy, compositional and electrophoresis studies confirmed the encapsulation episode of the biomaterial. In order to elucidate the use of this resulting nanobiocomposite as a delivery vector, an MTT assay was performed to determine any cytotoxic effects of the host towards cells. The intercalated pEGFP-N2 anion was later successfully recovered through acidification with HNO(3) after treatment with DNA-degrading enzymes, thus also showing the ability of the LDH host to protect the intercalated biomaterial from degradation. Cell transfection studies on Vero3 cells were then performed, where cells transfected with the nanobiocomposite exhibited fluorescence as early as 12 h post-treatment compared to naked delivery of the plasmid itself.
Fulltext Association of ABO blood groups with diabetes mellitus

Kamil M, Al-Jamal HA, Yusoff NM

Libyan J Med, 2010 Feb 08;5.
PMID: 21483592 DOI: 10.3402/ljm.v5i0.4847

So far no studies have been performed in Malaysia to look at association of diabetes mellitus (DM) with blood groups. We studied the association of ABO blood groups with DM type 2.
Newcastle disease virus infection promotes Bax redistribution to mitochondria and cell death in HeLa cells

Molouki A, Hsu YT, Jahanshiri F, Rosli R, Yusoff K

Intervirology, 2010;53(2):87-94.
PMID: 19955813 DOI: 10.1159/000264198

Newcastle disease virus (NDV) is an avian paramyxovirus that has gained a lot of interest in cancer viro-therapeutic applications because of its ability to selectively induce apoptosis in human cancer cells. However, the underlying mechanisms by which NDV induces apoptosis in human cancer cells are still not entirely understood.
Fulltext Filgrastim and antibiotics treatment reduces neutropenia severity in solid cancer patients

Hassan BA, Yusoff ZB, Othman SB

Asian Pac J Cancer Prev, 2009 Oct-Dec;10(4):641-4.
PMID: 19827886

INTRODUCTION: Neutropenia has a detrimental effect on cancer patients' quality of life, also possibly resulting in a reduction in the chemotherapy dose which could lead to an increment in the size of a cancer. The main danger associated with neutropenia is the risk of bacterial, fungal or viral infection, which may lead to patient death. Treatment including granulocyte-colony stimulating factors (G-CSF, filgrastim) so as to increase the body immunity is given to neutropenic patients with no infection i.e., absence of fever. However, when infection is present, antibiotics such as ceftazidime, imipenem and vancomycin need to be used.

OBJECTIVE: The aim of this study was to find the association between neutropenia severity and treatment with filgrastim (Neupogen) alone or in combination with antibiotics in solid cancer patients.

METHODS: This is an observational retrospective study on 117 cases suffering from neutropenia after chemotherapy administration. The patients were admitted to a government hospital for cancer treatment between the years 2003-2006. The types of data collected were categorical and not normally distributed, covering demography, chemotherapy, severity of neutropenia (classified on absolute neutrophil count into mild, moderate and severe) and treatment of neutropenia, either filgrastim (Neupogen) alone or in combination with antibiotics. Statistical tests used were the Chi-square test, Fisher's exact test and logistic regression.

RESULTS: The majority (69.2%) of the patients were treated with filgrastim (81) alone, only 30.8% receiving the combination. Significant associations between both treatments and neutropenia severity. Both Chi-square and Fisher's exact tests showed P= 0.001. Logistic regression showed that filgrastim is the major treatment for severe neutropenic patients since the result showed an infinity (E) and P= 0.001 for filgrastim alone more than its combination with antibiotic.

CONCLUSION: The use of filgrastim is highly associated with treatment of severe neutropenia in solid cancer patients who received chemotherapy. So filgrastim is considered as the drug of choice in the presence of severe neutropenic cases.
Fulltext 3-Oxoolean-1-en-28-oic acid-n-hexa-ne-water (4/1/1) from the bark of Walsura pinnata Hassk

Awang K, Yusoff M, Mohamad K, Chong SL, Ng SW

Acta Crystallogr Sect E Struct Rep Online, 2009;65(Pt 5):o1166.
PMID: 21583968 DOI: 10.1107/S1600536809015086

3-Oxoolean-1-en-28-oic acid, isolated from the bark of Walsura pinnata Hassk, crystallized from n-hexane as an n-hexane 0.25-solvent 0.25-hydrate, C(30)H(46)O(3)·0.25C(6)H(14)·0.25H(2)O. There are two independent mol-ecules in the asymmetric unit of the title compound. The three six-membered cyclo-hexane rings in each mol-ecule adopt chair conformations and the carboxyl substituent occupies an axial/equatorial position. The two independent mol-ecules are linked by a pair of O-H(carbox-yl)⋯O hydrogen bonds into a dimer. The n-hexane mol-ecule is disordered about a twofold rotation axis and the water mol-ecule lies on a twofold rotation axis. In addition, the cyclo-hexone carbonyl group of one of the independent mol-ecules is disordered over two sites with occupancies of 0.75 and 0.25.
Fulltext Pinnatane A from the bark of Walsura pinnata Hassk

Mohamad K, Yusoff M, Awang K, Ahmad K, Ng SW

Acta Crystallogr Sect E Struct Rep Online, 2009;65(Pt 6):o1317.
PMID: 21583174 DOI: 10.1107/S1600536809015955

In the mol-ecule of pinnatane A, C(30)H(48)O(3), isolated from the bark of Walsura pinnata Hassk, the four cyclo-hexane rings adopt chair conformations; the carboxyl and hydr-oxy substituents occupy axial positions. The cyclo-hexene ring is envelope-shaped. Adjacent mol-ecules are linked by O-H⋯O hydrogen bonds into a chain running along the c axis.
Interactions of hepatitis B core antigen and peptide inhibitors

Tang KF, Abdullah MP, Yusoff K, Tan WS

J Med Chem, 2007 Nov 15;50(23):5620-6.
PMID: 17918821

The core protein (HBcAg) of hepatitis B virus (HBV) has been shown to interact with the large surface antigen during HBV morphogenesis, and these interactions can be blocked by small peptides selected from either linear or constrained phage display peptide libraries. The association of HBcAg with peptide inhibitors was quantitatively evaluated by isothermal titration calorimetry. The thermodynamic data show that the interaction between HBcAg and peptide MHRSLLGRMKGA is enthalpy-driven and occurs at a 3:1 stoichiometry and dissociation constant (Kd) value of 79.4 muM. However, peptide WSFFSNI displays a higher binding affinity for HBcAg with a Kd value of 18.5 muM when compared to peptide MHRSLLGRMKGA. A combinatorial approach using chemical cross-linking and surface-enhanced laser desorption/ionization-time-of-flight-mass spectrometry shows that the Lys of peptide MHRSLLGRMKGA interacted either with D64, E77, or D78 of HBcAg.
A phage-displayed single chain variable fragment that interacts with hepatitis B core antigen: library construction, selection and diagnosis

Tan GH, Yusoff K, Seow HF, Tan WS

J Clin Virol, 2007 Jan;38(1):49-56.
PMID: 17074533

Phage display is an alternative method for constructing and selecting antibodies with desired specificity towards an antigen.
Recombinant Newcastle Disease virus capsids displaying enterovirus 71 VP1 fragment induce a strong immune response in rabbits

Sivasamugham LA, Cardosa MJ, Tan WS, Yusoff K

J Med Virol, 2006 Aug;78(8):1096-104.
PMID: 16789020

The complete VP1 protein of EV71 was truncated into six segments and fused to the C-terminal ends of full-length nucleocapsid protein (NPfl) and truncated NP (NPt; lacks 20% amino acid residues from its C-terminal end) of newcastle disease virus (NDV). Western blot analysis using anti-VP1 rabbit serum showed that the N-terminal region of the VP1 protein contains a major antigenic region. The recombinant proteins carrying the truncated VP1 protein, VP1(1-100), were expressed most efficiently in Escherichia coli as determined by Western blot analysis. Electron microscopic analysis of the purified recombinant protein, NPt-VP(1-100) revealed that it predominantly self-assembled into intact ring-like structures whereas NPfl-VP(1-100) recombinant proteins showed disrupted ring-like formations. Rabbits immunized with the purified NPt-VP(1-100) and NPfl-VP(1-100) exhibited a strong immune response against the complete VP1 protein. The antisera of these recombinant proteins also reacted positively with authentic enterovirus 71 and the closely related Coxsackievirus A16 when analyzed by an immunofluorescence assay suggesting their potential as immunological reagents for the detection of anti-enterovirus 71 antibodies in serum samples.
Fulltext Generalized epilepsy with febrile seizure plus (GEFS+) spectrum: Novel de novo mutation of SCN1A detected in a Malaysian patient

Tan EH, Yusoff AA, Abdullah JM, Razak SA

J Pediatr Neurosci, 2012 May;7(2):123-5.
PMID: 23248692 DOI: 10.4103/1817-1745.102575

In this report, we describe a 15-year-old Malaysian male patient with a de novo SCN1A mutation who experienced prolonged febrile seizures after his first seizure at 6 months of age. This boy had generalized tonic clonic seizure (GTCS) which occurred with and without fever. Sequencing analysis of voltage-gated sodium channel a1-subunit gene, SCN1A, confirmed a homozygous A to G change at nucleotide 5197 (c.5197A > G) in exon 26 resulting in amino acid substitution of asparagines to aspartate at codon 1733 of sodium channel. The mutation identified in this patient is located in the pore-forming loop of SCN1A and this case report suggests missense mutation in pore-forming loop causes generalized epilepsy with febrile seizure plus (GEFS+) with clinically more severe neurologic phenotype including intellectual disabilities (mental retardation and autism features) and neuropsychiatric disease (anxiety disorder).

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